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Fetal programming

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Ontogeny*

Phylogeny*

Developmental Plasticity

Devo � Evo

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Mismatch/DOHA6

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XXI Century Epidemiologic

Transition

Environment

From Genetics to Epigenetics

Evolutionary Medicine

ERNESTO BURGIO

ECERI - European Cancer and Environment Research InstituteISDE Scientific Committee

Electromagnetic Fields Fetal Effects

Seven keys towards a paradigm shift in biomedicine:environmental interference with the human (epi)geno me

We are currently facing a paradigm shift in biomedicine

For the last 50 years it was agreed to consider DNA as the code

and the key project for the assembly of our phenotype.

In the last ten years and especially since the appearance of the first molecular epigenetic studies we have begun to understand

that the construction of the phenotype is the result of the interaction between the information coming from the environment

and the information deeply inscribed inside the DNA

thanks to a very complex molecular network surrounding DNA: the epigenome

Therefore it can be argued that there is no stable change in our phenotype (both physiological and pathological) which is not

- environmentally induced

- modulated by the epigenome

- conditioned by the DNA

Other key concepts (obviously interdependent) are:

- developmental plasticity

- fetal programming

allow ing us to understand how the fetus epigenetically program (for life) all its cells in a predictive and adaptive way

responding to information coming from the environment (through the mother bias )

It is important to note that in this period

incorrect information ( pollutants, endocrine disruptors ..) and /or

discrepancies between the information that the baby receives before and after birth (mismatch)

may create epigenetically bad programmed cells (including gametes), thus causing chronic diseases in adulthood or even in subsequent generations

This theory (DOHaD Developmental Origins of Health and Disease)

could help us to explain the current epidemiological transition ..

Obesity/Metabolic Syndrome/Diabetes 2

Neurobehavioral Deficits

and DiseasesCANCER

Reproductive

Diseases/Dysfunctions

Multiorgan Effects of

Endocrine Disruptors

In Vitro Fertilization

Semen Abnormalities

Obesogens

DOHAD

Asthma and allergies

Psychiatric Diseases

Cardiovascular

Diseases

Ipertension

Developmental Time Windows of

Vulnerability

Pesticides

Lung Development

Placenta: Prediction

of Future Health

Materno Fetal Stress

Eventually, during the last years, the fetal programming mismatch theoryhas been transformed from a theory essentially useful to explain the pathogenic mechanisms causing certain diseases of adulthood,

into the key-model theory of the embryo-fetal origin s of adult diseases (DOHA-Developmental Origins of Health and Diseases)

Interphase chromosome

Mitotic chromosome

Euchromatin

Heterochromatin

Our proposal is not to consider epigenetics just as a (minor) part of genetics (the marks of DNA, the histone modifications, the interference of microRNAs modulating in various ways the transcription and translation of the message contained in the DNA), but to recognize in the study of epigenetics the most appropriate and powerful tool to build up a new systemic and molecular model of genome , finally understood as a dynamic and fluid network which can interact insid e itself and with the outside

The first keyword: Epigenetics

Rudolf Jaenisch- Whitehead Institute and

Dept. of Biology, MIT, Cambridge, MA

In such a fluid and systemic model the epigenome (also defined by some scientists as the controlling software of the genome) behaves as a sort of compensation chamber - the specific place where the flow of information that comes from outside (environment and microenvironment) meets and interacts with the information encoded in the genes for millions years (the hardware )

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http://news.sciencemag.org/sciencenow/2009/04/21-03.html IN FACT Genes need to be told

to switch “off” and “on”:• Genes need to be told how much expression

(protein) is required and where.

• Genes need to be regulated – this regulation is not

performed by DNA but by many other controls

arranged in a complex network

• DNA has been called the Book of Life by the Human

Genome Project scientists, but many other biologists

consider DNA to be simply a random collection of

words from which a meaningful story of life may be

assembled…

• In order to assemble that meaningful story, a living

cell uses a second informational system. (...) The key

concept here is that these dynamic-epigenetic

networks have a life of their own —they

follow network-rules not specified by DNA

From directing the fate of stem cells to determining how.. we grow, the genes in our body

act in complex networks.. the whole Genome is a Complex and highly dynamic molecular

Network of interacting Genes and non-codifying sequences.. and proteins

Strohman R. , April 2001 Beyond genetic determinism ….Genes Know How to Network…BUT...

TCDD

Viruses

HERVs

EMF

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SYNERGISM !!

“FLUID EPI-GENOME”

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We may represent the environment as a continuous

stream of information (simple: photons: individual

packages of E = M = Information) or complex (organic

molecules, viruses etc) interacting with our cells

[membrane /transmembrane receptors, signal

transduction proteins, nuclear receptors, genome

(DNA + Epigenome)] forcing them to adapt

The second keyword: Environment

Electromagnetic fields (EMF), in both ELF (extremely low

frequency) and radio frequency (RF) ranges, activate the

cellular stress response, a protective mechanism

that induces the expression of stress response genes, e.g.,

HSP70, and increased levels of stress proteins hsp70…

(The 20 different stress protein families are evolutionarily

conserved and act as ‘chaperones’ in the cell when they

‘help’ repair and refold damaged proteins and transport

them across cell membranes..)

Induction of the stress response involves activation

of DNA, and despite the large difference in energy

between ELF and RF, the same cellular pathways

respond in both frequency ranges.

Specific DNA sequences on the promoter of the

HSP70 stress gene are responsive to EMF..

While low energy EMF interacts with DNA to induce the stress response,

increasing EMF energy in the RF range can lead to breaks in DNA strands.

PathophysiologyVolume 16, Issues 2-3, August 2009, Pages 71-78

Specific DNA sequences on the promoter of the HSP70 stress gene are responsive to EMF...

Hsf-1

Synthesis of this stress protein is initiated in a region of the promoter where a transcription factor known as Heat Shock Factor 1 (HSF-1) binds to a Heat Shock E lement (HSE).The EMF sensitive region on HSP70 promoter is upstream from the thermal domain of the promoter and is not sensitive to increased temperat ure . The binding of HSF-1 to HSE occurs at −192 in the HSP70 promoter relative to the transcription initiation site.

The EMF domain contains three nCTCTn myc-binding sites −230, −166 and −160 relative to the transcription initiation site and upstream of t he binding sites for the heat shock (nGAAn ) and serum responsive elements…. The electromagnetic response elements (EMREs) have also been identified on the c- myc promoter and are also responsive to EMF

HSP70 stress gene

PathophysiologyVolume 16, Issues 2-3, August 2009, Pages 71-78

… a technical term that refers to the capability and, at the same time, the requirement, for embryo-foetal cells to define their epigenetic setting in a predictive and adaptive way , in relation to the information coming from the mother and, through her, from the outer world ..

A predictive adaptive response (PAR) is a developmental trajectory taken by an organism during a period of developmental plasticity in response to perceived environmental cues..

Dioxin and Dioxin-like molecules

(Ultra)-fine particles

Heavy Metals

Polycyclic Aromatic Hydrocarbons (PAH)

Benzene

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The third key word is fetal programming …

MATERNAL

STRESS

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Fetal Programming

Differentiation

epi-mutations

Cellular Differentiation: an epigentic process Differentiation occurs numerous times

during the development of a multicellular

organism as the organism changes from

a single zygote to a complex system of

tissues and 200 cell types (genetically

identical.. each with its own epigenetic

and morpho-functional characteristics)..

Gametogenesis. Maturation of germ cells is characterized by

an impressive degree of cellular restructuring and gene

regulation that involves remarkable genomic reorganization.

These events are finely tuned, but are also susceptible to the

introduction of various types of error…

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Nature 447, 425-432 (24 May 2007)

Developmental

PLASTICITY

The fourth keyword is developmental plasticity

In contrast to what it is often asserted, DNA does not contain the genetic

program of the individual, but simply an enormous amount of potential

information.. Developmental plasticity refers to the many possible

phenotypes (polyphenisms) that may result from a single genome.

Belyaev et al [2010] reported that 915 MHz microwave exposure significantly affects human stem cells

“The strongest microwave effects were always observed in stem cells. This result may suggest both significant misbalance in DSB repair, and severe stress response.

Our findings that stem cells are the most sensitive to microwave exposure, and react to more frequencies than do differentiated cells may be important for cancer risk assessment and indicate that

stem cells are the most relevant cellular model for validating safe mobile communication signals.”

Belyaev I, Markova E, Malmgren L. [2010] Microwaves from Mobile Phones Inhibit 53BP1 Focus

Formation in Human Stem Cells Stronger than in Differentiated Cells: Possible Mechanistic Link

to Cancer Risk. Environ Health Perspect. 118(3): 394–399

Chen C, Ma Q, Liu C, Deng P, Zhu G, Zhang L, He M, Lu Y, Duan W, Pei L, Li M, Yu Z,

Zhou Z Exposure to 1800 MHz radiofrequency radiation impai rs neuriteoutgrowth of Embryonic neural stem cells . Sci Rep. 2014 May 29;4:5103A radiofrequency electromagnetic field (RF-EMF) of 1800 MHz is widely used in mobile

communications. However, the effects of RF-EMFs on cell biology are unclear. Embryonic neural stem

cells (eNSCs) play a critical role in brain development. Thus, detecting the effects of RF-EMF on eNSCs is

important for exploring the effects of RF-EMF on brain development. We exposed eNSCs to 1800 MHz

RF-EMF at specific absorption rate (SAR) values of 1, 2, and 4 W/kg for 1, 2, and 3 days. We found that

1800 MHz RF-EMF exposure did not influence eNSC apoptosis, proliferation, cell cycle or the mRNA

expressions of related genes. RF-EMF exposure also did not alter the ratio of eNSC differentiated neurons

and astrocytes. However, neurite outgrowth of eNSC differentiated neurons was inhibited after

4 W/kg RF-EMF exposure for 3 days. Additionally, the mRNA and protein expression of the

proneural genes Ngn1 and NeuroD, which are crucial for neurite outgrowth, were decreased

after RF-EMF exposure. The expression of their inhibitor Hes1 was upregulated by RF-EMF

exposure. These results together suggested that

1800 MHz RF-EMF exposure impairs neurite

outgrowth of eNSCs. More attention should

be given to the potential adverse effects of

RF-EMF exposure on brain development.

Disturbing the CONNECTOME INSTRUCTION

Ma Q, Deng P, Zhu G, Liu C, Zhang L, Zhou Z, Luo X, Li M, Zhong M, Yu Z, Chen C, Zhang Y

Extremely low-frequency electromagnetic fields affe ct transcript levels ofNeuronal differentiation-related genes in embryonic neural stem cells . PLoS One

2014 Mar 3;9(3):e90041. doi: 10.1371/journal.pone.0090041. eCollection 2014.

Previous studies have reported that extremely low-frequency electromagnetic fields (ELF-EMF) can affect

the processes of brain development, but the underlying mechanism is largely unknown. The proliferation

and differentiation of embryonic neural stem cells (eNSCs) is essential for brain development during the

gestation period. To date, there is no report about the effects of ELF-EMF on eNSCs. In this paper, we

studied the effects of ELF-EMF on the proliferation and differentiation of eNSCs. Primary cultured eNSCs

were treated with 50 Hz ELF EMF; various magnetic intensities and exposure times were applied.

Our data showed that there was no significant change in cell proliferation, which was evaluated by cell

viability (CCK-8 assay), DNA synthesis (Edu incorporation), average diameter of neurospheres, cell cycle

distribution (flow cytometry) and transcript levels of cell cycle related genes (P53, P21 and GADD45

detected by real-time PCR). When eNSCs were induced to differentiation, real-time PCR results showed a

down regulation of Sox2 and up-regulation of Math1, Math3, Ngn1 and Tuj1 mRNA levels after 50 Hz ELF

EMF exposure (2 mT for 3 days), but the percentages of neurons (Tuj1 positive cells) and astrocytes (GFAP

positive cells) were not altered when detected by immunofluorescence assay.

Although cell proliferation and the percentages of neurons and astrocytes

differentiated from eNSCs were not affected by 50 Hz ELF-EMF, the expression

of genes regulating neuronal differentiation was altered.

In conclusion, our results support that 50 Hz ELF-EMF induce molecular

changes during eNSCs differentiation, which might be compensated

by post-transcriptional mechanisms to support cellular homeostasis.

Disturbing the CONNECTOME INSTRUCTION

Early critical periods in the development

of SYNAPTOGENESIS and brain functions

Formation of new synapses following stimulation..

The Individual wiring

Disturbing the CONNECTOME INSTRUCTION

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Brain plasticity and modulation of its structure

and its functions

Motility of neurons and in particular the formation of new connections (synapses) can be modified (perturbed) by exposure to environmental stressors

Wingate Imagining the brain cell: the neuron in visual culture. Nature Rev Neuroscience 2006; 7: 745-752.

The Individual wiringDisturbing the CONNECTOME INSTRUCTION

The brain grows at an amazing rate during

development. At times during brain

development, 250,000 neurons are

added every minute!

At birth, almost all the neurons that the

brain will ever have are present.

However, the brain continues to grow for

many years after birth.

By the age of 2 years old, the brain is

about 80% of the adult size

A stegosaurus dinosaur weighed approximately 1,600 kg but had a brain that weighed only

approximately 70 grams (0.07 kg). Therefore, the brain was only 0.004% of its total body

weight. In contrast, an adult human weighs approximately 70 kg and has a brain that weighs

approximately 1.4 kg. Therefore, the human brain is about 2% of the total body weight. This

makes the brain to body ratio of the human 500 times greater than that of the stegosaurus

Disturbing the CONNECTOME INSTRUCTION

The gift our mothers

never wanted to give us

http://www.ewg.org/reports/generations/

CHEMICAL FALL OUT

ULTRAFINE PARTICLESHEAVY METALS

ENDOCRINE DISRUPTORS

dioxin-like moleculles1

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.. at present many studies in various parts of the world are evaluating the chemical body burden .. especially in women, children, embryos / fetuses, providing dramatic results .

Giuseppe Giordano

Monitoring Body-Burdens

700 different synthetic chemicals or heavy

metals found in human blood,

POPs

..a growing overload of electromagnetic radiations is adding to chemical toxicburden: here we demonstrate that the fetal exposure to 800–1900 Mhz-rated radio-frequency radiation from cellular telephones leads to behavioral and neurophysiological alterations that persist into ad ulthood .

Mice exposed during pregnancy hadimpaired memory, werehyperactive, and had increasinganxiety, indicating that in-uteroexposure to radiofrequencyis a potential cause ofneurobehavioral disorders.

• We further demonstratedimpairment of glutamatergicsynaptic transmission ontopyramidal cells in the prefrontalcortex associated with thesebehavioral changes

• suggesting a mechanism by whichin-utero cellular telephoneradiation exposure may lead tothe increased prevalenceof neurobehavioral disorders.